Analysis of addiction antitoxin MazE by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0799-WXH)
Introduction
The Escherichia coli mazEF addiction module plays a crucial role in the cell death program that is triggered under various stress conditions. It codes for the toxin MazF and the antitoxin MazE, which interferes with the lethal action of the toxin.
Differential Scanning Fluorimetry measures protein thermal unfolding by monitoring changes in fluorescence emission of a sample upon heating. This allows the determination of protein thermostability and complex formation even with weakly binding ligands by thermal shift assay. Differential Scanning Fluorimetry is therefore ideally suited for screening of optimum buffer conditions like pH, buffer composition and ionic strength. The technique is applicable to any biological sample, from soluble proteins to integral membrane proteins.
Applications
To identify low-molecular-weight ligands that bind and stabilize purified proteins. To measure the denaturation and unfolding of proteins.
Procedure
1. Preparation of compound solutions 2. Preparation of buffer/additive screen plates 3. Preparation of compound storage plates 4. Equipment preparation 5. Sample preparation 53. Performing the scan